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The zone between plaque and attached periodontal tissues on chronic periodontitis-affected teeth was examined by a combined macroscopic and scanning electron microscopy (SEM) approach. Examined were 27 teeth with chronic periodontitis (chronic periodontitis-affected group) and three healthy teeth with no evidence of periodontal disease (control group). Both groups were collected immediately after extraction, fixed in 2% glutaraldehyde, and post-fixed in 1% osmium tetroxide. Then, teeth were macroscopically examined to identify their stained zones. Teeth were dehydrated, critical-point dried, gold coated, and examined in an SEM. Both healthy and chronic periodontitis-affected teeth showed a very similar staining pattern on their surfaces. An unstained band with a belt-like appearance was observed around the teeth, delimited by two osmium-stained zones. Some weakly stained areas were frequently observed inside the unstained band. The SEM examination showed four different regions in both groups. These regions appeared in the following coronoapical sequence: dental plaque, plaque-free zone, junctional epithelium, and attached periodontal tissues. A dental cuticle covering the cementum surface from the plaque border to the junctional epithelium was detected on chronic periodontitis-affected teeth. Some aspects of this particular zone may be involved in the pathogenesis of inflammatory periodontal disease and therefore may have some influence on treatment for chronic periodontitis-affected teeth. [ABSTRACT FROM AUTHOR]

OBJECTIVES: The aim of this study was to ultrastructurally examine the influence of simvastatin on bone healing in surgically created defects in rat mandibles. STUDY DESIGN: Bone defects 0.8 mm in diameter were created in the buccal aspect of first mandibular molar roots and filled with 2.5% simvastatin gel, while the controls were allowed to heal spontaneously. The rats were humanely killed 7, 9, 11, or 14 days postoperatively, and the specimens were processed for scanning and transmission electron microscopy, as well as for colloidal gold immunolabeling of osteopontin. RESULTS: The regenerated alveolar bone in the simvastatin-treated defects presented smaller marrow spaces, and the collagen fibrils were regularly packed exhibiting a lamellar bone aspect. Osteopontin was present through the bone matrix during the wound healing and alveolar bone regeneration. CONCLUSION: The present study provides evidence that a single topical application of 2.5% simvastatin gel improves the quality of the new bone and decreases bone resorption. [ABSTRACT FROM AUTHOR]

An ultrastructural study of the cementum and periodontal ligament (PDL) changes after continuous intrusion with two different and controlled forces in humans was carried out. Twelve first upper premolars, at stage 10 of Nolla, orthodontically indicated for extraction from six patients (mean age 15.3) were used. They were divided into three experimental groups, distributed intra-individually as follows: control (not moved), continuously intruded for 4 weeks with 50 or 100 cN force, utilizing a precise biomechanical model with nickel titanium super-elastic wires (NiTi-SE), which were developed and calibrated individually. The teeth were extracted, fixed, decalcified, and conventionally processed for examination in a Jeol 100 CX II transmission electron microscope. Evident signs of degeneration of cell structures, vascular components, and extracellular matrix (EM) of cementum and PDL were observed in all the intruded teeth, with more severe changes towards an apical direction and in proportion to the magnitude of force applied. Resorptive areas and an irregular root surface of the intruded teeth were noticed, according to the same pattern described above. Concomitant, areas of repair were also revealed in the cementum and PDL although the magnitude of forces remained the same throughout the experimental period. Thus, a reduction of continuous force magnitude should be considered to preserve the integrity of tissues. [ABSTRACT FROM AUTHOR]

In recent years, alternative pulpal therapies targeting dentinogenesis signaling pathways using different peptides have been investigated. The aim of this study was to verify the effectiveness of poly(aspartic acid), pAsp, in dentin regeneration using an animal model., Methods: Mechanical pulp exposure was performed in the upper molars of 56 Wistar rats, randomly divided as follows (n = 14): control (no treatment); MTA group-pulp capping with mineral trioxide aggregate (MTA Angelus); pAsp group-application of 20 μL of pAsp solution (25 mg·mL -1 ); MTA+pAsp group-application of MTA mixed with pAsp (5:1 by mass). Animals were euthanized after 7 or 21 days. Histological sections were submitted to hematoxylin-eosin and Brown and Brenn staining and immunohistochemical analysis for osteopontin (OPN) and dentin matrix protein 1 (DMP 1)., Results: At 7 days, an acute inflammatory infiltrate and the presence of disorganized mineralized tissue were observed in all groups. At 21 days, the quality and thickness of the reparative dentin in treated groups were superior to the control, and bacterial contamination was observed in two MTA-pAsp specimens. While all treated groups showed intense immunostaining for OPN at 21 days, only the pAsp group expressed DMP 1, indicating the presence of fully differentiated odontoblast-like cells., Conclusion: Poly(aspartic) acid promoted dentin regeneration in rat molars in the absence of an additional calcium source and may be an alternative to MTA as a pulp-capping agent.

The association of more than one wavelength for photobiomodulation therapy (PBMT) to treat oral mucositis (OM) is unusual in the literature. Thus, this study aims to compare the simultaneous irradiation effects with their isolated application to treat OM. In order of that, 48 male Syrian hamsters were divided into 4 groups: Chemotherapy (Ch), which received only a OM induction protocol (5-fluorouracil chemotherapy and superficial oral mucosa scratches); red laser (RL), which received the OM induction and a PBMT protocol at 660 nm; infrared laser (IRL), which received the OM induction, and a PBMT protocol at 808 nm; and the RL + IRL group, which received the simultaneous application, of 660 and 808 nm wavelengths. Clinical (OM grade classification), histological (light microscopy analysis with H&E and collagen staining), immunohistochemical (TNF-α expression), and biochemical (TNF-α and hydroxyproline concentration) analyzes were performed after 7 and 10 days. Mainly on the 10th day, the RL and IRL groups showed lower OM grades and faster microscopic repair process, with greater expression of collagen fibers and lower TNF-α levels, besides the higher hydroxyproline concentrations, mainly in comparison with the Ch group. In conclusion, in this study, the simultaneous protocol did not present superior results than the isolated irradiations., (© 2023 Wiley-VCH GmbH.)

The unbridled dissemination of multidrug-resistant pathogens is a major threat to global health and urgently demands novel therapeutic alternatives. Antimicrobial photodynamic therapy (aPDT) has been developed as a promising approach to treat localized infections regardless of drug resistance profile or taxonomy. Even though this technique has been known for more than a century, discussions and speculations regarding the biochemical mechanisms of microbial inactivation have never reached a consensus on what is the primary cause of cell death. Since photochemically generated oxidants promote ubiquitous reactions with various biomolecules, researchers simply assumed that all cellular structures are equally damaged. In this study, biochemical, molecular, biological and advanced microscopy techniques were employed to investigate whether protein, membrane or DNA damage correlates better with dose-dependent microbial inactivation kinetics. We showed that although mild membrane permeabilization and late DNA damage occur, no correlation with inactivation kinetics was found. On the other hand, protein degradation was analyzed by three different methods and showed a dose-dependent trend that matches microbial inactivation kinetics. Our results provide a deeper mechanistic understanding of aPDT that can guide the scientific community toward the development of optimized photosensitizing drugs and also rationally propose synergistic combinations with antimicrobial chemotherapy., (© 2022 American Society for Photobiology.)

Antimicrobial blue light (aBL) offers efficacy and safety in treating infections. However, the bacterial targets for aBL are still poorly understood and may be dependent on bacterial species. Here, we investigated the biological targets of bacterial killing by aBL (λ = 410 nm) on three pathogens: Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa. Initially, we evaluated the killing kinetics of bacteria exposed to aBL and used this information to calculate the lethal doses (LD) responsible for killing 90 and 99.9% of bacteria. We also quantified endogenous porphyrins and assessed their spatial distribution. We then quantified and suppressed reactive oxygen species (ROS) production in bacteria to investigate their role in bacterial killing by aBL. We also assessed aBL-induced DNA damage, protein carbonylation, lipid peroxidation, and membrane permeability in bacteria. Our data showed that P. aeruginosa was more susceptible to aBL (LD 99.9 = 54.7 J/cm 2 ) relative to S. aureus (LD 99.9 = 158.9 J/cm 2 ) and E. coli (LD 99.9 = 195 J/cm 2 ). P. aeruginosa exhibited the highest concentration of endogenous porphyrins and level of ROS production relative to the other species. However, unlike other species, DNA degradation was not observed in P. aeruginosa. Sublethal doses of blue light (90 ) could damage the cell membrane in Gram-negative species but not in S. aureus. In all bacteria, oxidative damage to bacterial DNA (except P. aeruginosa), proteins, and lipids occurred after high aBL exposures (>LD 99.9 ). We conclude that the primary targets of aBL depend on the species, which are probably driven by variable antioxidant and DNA-repair mechanisms. IMPORTANCE Antimicrobial-drug development is facing increased scrutiny following the worldwide antibiotic crisis. Scientists across the world have recognized the urgent need for new antimicrobial therapies. In this sense, antimicrobial blue light (aBL) is a promising option due to its antimicrobial properties. Although aBL can damage different cell structures, the targets responsible for bacterial inactivation have still not been completely established and require further exploration. In our study, we conducted a thorough investigation to identify the possible aBL targets and gain insights into the bactericidal effects of aBL on three relevant pathogens: Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa. This research not only adds new content to blue light studies but opens new perspectives to antimicrobial applications.

Osteoclasts are specialized cells that degrade and resorb bone. Bisphosphonates (BPs) are drugs with well-known capacity to inhibit the resorption of mineralized tissues. Nitrogen-containing BPs, like alendronate (ALN) and zoledronic acid (ZA), inactivate osteoclast activity mostly by alterations on the cytoskeleton architecture of the cell. In this study, we used an in vitro model to test the hypothesis that bisphosphonates may have inhibitory effects on the osteoclastogenesis and osteoclast activity after the therapy was discontinued. Primary osteoclasts were generated from mouse bone marrow in media supplemented with 1,25-dihydroxyvitamin D3 and cultivated over bones pre-treated with ALN and ZA. The pre-saturation of the bone slices with bisphosphonates did not affect cell viability. We found, however, that by disrupting the gene expression of RANKL and OPG the osteoclastogenesis and resorption activity of osteoclasts was significantly disturbed. These inhibitory effects were confirmed by scanning electron microscopy resorption assay, assessment of osteoclast ultrastructure, and by gene expression analysis of TRAP and Cathepsin K. In conclusion, ALN and ZA adhered to the bone matrix reduced the osteoclast activity in vitro., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

Osteoclasts are cells whose main function is the resorption of bone matrix. However, several factors, including medications, can interfere with the resorption process. Alendronate (ALN), a nitrogen-containing type of bisphosphonate, and dexamethasone (DEX), a glucocorticoid, are drugs that may affect the resorption activity. The aim of this study is to investigate the effects of ALN, and/or DEX on osteoclast gene expression and resorption activity in primary mouse marrow cultures stimulated with 1,25-dihydroxyvitamin D3, a model for the bone microenvironment. Cultures were treated only with ALN (10 -5 M), DEX (10 -6 M), and with a combination of both agents. Viability assays performed at days 5, 7, and 9 showed the highest number of viable cells at day 7. All the following assays were then performed at day 7 of cell culture: tartrate resistant acid phosphatase (TRAP) histochemistry, receptor activator of nuclear factor kappa B ligand (RANKL) immunofluorescence, osteoprotegerin (OPG), and RANKL gene expression by qPCR and resorption analysis by scanning electron microscopy. Treatment with ALN, DEX, and the combination of both did not promote significant changes in the number of TRAP+ cells, although larger giant cells were detected in groups treated with DEX. DEX treatment increased the gene expression of RANKL and reduced OPG. The treatment with ALN reduced the depth of the resorption pits, but their inhibitory effect was less effective when administered with DEX.

Objectives: The aim of this study was to analyze the effects of three different Photobiomodulation Therapy (PBMT) protocols in the treatment of 5-fluorouracil-induced oral mucositis in hamsters., Design: 60 hamsters were divided into five groups: group "C", which did not receive oral mucosa scratching, 5-fluorouracil (5-FU) or PBMT; group "Ch", which received anesthesia, superficial oral mucosa scratching and 5-FU (oral mucositis induction); and three groups that received oral mucositis induction and a PBMT protocol: groups ChLI, ChLII and ChLIII that received 0.24 J (one point), 1 J (one point) and 1.2 J (five points of 0.24 J) of energy, respectively. The laser equipment used had λ = 660 nm and 0.04 cm 2 of spot area (0.226 cm diameter). The animals were euthanized on days 7 and 10 of the experiment, and their oral mucosas were removed for histological (light microscopy and collagen staining), immunohistochemical (NF-kB and TNF-α), and biochemical (TNF-α, NF-kB and hydroxyproline) analysis., Results: Group ChLI (less energy), showed the most accelerated repair rates and a lower concentration of inflammatory biomarkers than group Ch. Comparing the three PBMT protocols for treatment of 5-FU-induced oral mucositis in hamsters, the one with low energy (0.24 J) showed better results, regarding reduction of inflammatory biomarkers and tissue repair, than the ones with higher energy (1 and 1.2 J)., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Mesenchymal and epithelial stem cells were identified in dental tissues; however, knowledge about the odontogenic stem cells is limited, and there are some questions regarding their temporo-spatial dynamics in tooth development., Objective: Our study aimed to analyze the expression of the stem cell markers CD146 and p75NTR during the different stages of odontogenesis., Methodology: The groups consisted of 13.5, 15.5, 17.5 days old embryos, and 14 days postnatal BALB/c mice. The expression of CD146 and p75NTR was evaluated by immunohistochemistry., Results: Our results showed that positive cells for both markers were present in all stages of tooth development, and the number of positive cells increased with the progression of this process. Cells of epithelial and ectomesenchymal origin were positive for CD146, and the expression of p75NTR was mainly detected in the dental papilla and dental follicle. In the postnatal group, dental pulp cells were positive for CD146, and the reduced enamel epithelium and the oral mucosa epithelium showed immunostaining for p75NTR., Conclusions: These results suggest that the staining pattern of CD146 and p75NTR underwent temporal and spatial changes during odontogenesis and both markers were expressed by epithelial and mesenchymal cell types, which is relevant due to the significance of the epithelial-ectomesenchymal interactions in tooth development.

Objective: An ideal scaffold for endodontic regeneration should allow the predictableness of the new tissue organization and limit the negative impact of residual bacteria. Therefore, composition and functionalization of the scaffold play an important role in tissue bioengineering. The objective of this study was to assess the morphological, physicochemical, biological and antimicrobial properties of a new solid chitosan-based scaffold associated with gelatin, microparticulate dentin and genipin., Methods: Scaffolds based on chitosan (Ch); chitosan associated with gelatin and genipin (ChGG); and chitosan associated with gelatin, microparticulate dentin and genipin (ChGDG) were prepared by using the freeze-drying method. The morphology of the scaffolds was analyzed by scanning electron microscopy (SEM). The physicochemical properties were assessed for biodegradation, swelling and total released proteins. The biological aspects of the scaffolds were assessed using human cells from the apical papilla (hCAPs). Cell morphology and adhesion to the scaffolds were evaluated by SEM, cytotoxicity and cell proliferation by MTT reduction-assay. Cell differentiation in scaffolds was assessed by using alizarin red assay. The antimicrobial effect of the scaffolds was evaluated by using the bacterial culture method, and bacterial adhesion to the scaffolds was observed by SEM., Results: All the scaffolds presented porous structures. The ChCDG had more protein release, adhesion, proliferation and differentiation of hCAPs, and bacteriostatic effect on Enterococcus faecalis than Ch and ChGG (p < 0.05)., Significance: The chitosan associated with gelatin, microparticulate dentin and genipin has morphological, physicochemical, biological and antibacterial characteristics suitable for their potential use as scaffold in regenerative endodontics., (Copyright © 2021 The Academy of Dental Materials. Published by Elsevier Inc. All rights reserved.)

Objective: The aim of this study was to analyze the microhardness of demineralized enamel following different treatments (fluoride varnish, Er:YAG laser, and Er:YAG laser associated with fluoride varnish). Methods: Forty-eight enamel blocks (4 × 4 × 7 mm) were divided into six groups ( n  = 8): (S) Sound; (DE) Demineralized; (DED) DE + Duraphat ® 5% (fluoride varnish); (DEL20) DE + Er:YAG laser (20 mJ pulse mode; 0.20 W; 10 Hz; 60 sec; 1.18 J/cm 2 ; 11.83 W/cm 2 ); (DEL50) DE + Er:YAG laser (50 mJ pulse mode; 0.50 W; 10 Hz; 60 sec; 2.95 J/cm 2 ; 29.58 W/cm 2 ); (DEL20D) DE + Er:YAG laser (20 mJ) + Duraphat 5%. The irradiation was performed at 1 mm distance from the surface using a tip (AS7066X, L-14 mm, D-1.3 mm in diameter) in water/air spray refrigeration (level 6). The enamel blocks were submitted to pH cycling (4 h into DES solution +20 h into RE solution for 8 days and the solutions were changed every day). Knoop microhardness was measured (50 g/15 sec, six readings per sample) and data were analyzed by Kruskal-Wallis test at 5% significance. Results: After treatments, DF group showed higher microhardness values than all the groups. Also, DEL20D group showed similar results with H group according to the microhardness analysis ( p  < 0.05). Conclusions: It could be concluded that Duraphat 5% treatment showed better results when compared with all tested groups, however, the association of Er:YAG Laser 20 with Duraphat 5% also showed promising results.

The purpose of this study was to synthesize dicalcium phosphate dihydrate (DCPD) particles functionalized with triethylene glycol dimethacrylate (TEGDMA) through different routes by varying the receptor solution: ammonium phosphate (AP groups) or calcium nitrate (CN groups) and the moment in which TEGDMA was incorporated: ab initio (ab) or at the end of dripping the solution (ap). Two syntheses were performed without adding TEGDMA (nf). The particles were characterized by X-ray diffractometry, true density (using a helium pycnometer), surface area, and scanning electron microscopy. A 20 vol% of DCPD particles from the D, E, and F groups was added to the resin matrix to determine the degree of conversion (DC), biaxial flexural strength (BFS), the flexural modulus (FM), and surface roughness after an abrasive challenge (RA). A group with silanized barium glass particles was tested as a control. The data were submitted to ANOVA/Tukey's test (DC, BFS, and RA), and the Kruskal-Wallis test (FM) (alpha = 0.05). BFS values varied between 83 and 142 MPa, and the CN&#95;ab group presented a similar value (123 MPa) to the control group. FM values varied between 3.6 and 8.7 GPa (CN&#95;ab and CN&#95;nf groups, respectively), with a significant difference found only between these groups. RA did not result in significant differences. The use of calcium nitrate solution as a receptor, together with ab initio functionalization formed particles with larger surface areas. Higher BFS values were observed for the material containing DCPD particles with a higher surface area. In general, the DC, FM, and RA values were not affected by the variables studied.

Introduction: Oral mucositis (OM) has been considered one of the most feared collateral effects of oncological treatments. Some therapies have been used, such as light-emitting diode (LED), with promising results, but with no sufficient evidence in the literature. Objective: Our study aimed to evaluate, by clinical and histological analysis, the effect of LED on the treatment of chemotherapy-induced OM (CIOM) in an animal model. Methods: Twenty male hamsters were equally distributed to two groups: control (C), which received anesthesia and CIOM induction; and LED (L), which received anesthesia, CIOM induction, and LED treatment (635 nm, 120 mW, 0.48 J). The clinical analysis was performed through two specific scales for OM analysis on days 5, 7 and 10 of the experiment. In addition, the injured area of all hamsters check pouch mucosa was removed and processed for histological analysis on the last experimental day. Results: After statistical analysis, group L showed less severity of OM when compared with the C group ( P <0.05); beyond that, both healed completely on day 10. Conclusion: Our results suggested that the phototherapy with LED had a positive effect on accelerating repair, reducing the severity of CIOM., (Copyright © 2020 J Lasers Med Sci.)

Introduction: The primary aim of this randomized in vitro study was to compare the effectiveness of carbide, fibreglass and polymer burs on resinous remnant removal after bracket debonding, by the evaluation of enamel surface roughness and morphology. The secondary objective was to compare the time dispended on the procedures., Methods: The buccal surfaces of 28 bovine incisors were analysed by a profilometer to initial roughness measurement (Ra-T1). Brackets were bonded with a light-cured resin and debonded with a debonding plier. The samples were randomly divided into four groups, according to the bur used (n=7): A-Tungsten carbide; B-Fibreglass; C-Polymer; D-Polymer with 75% ethanol pre-treatment. The second roughness measurements were made after resin removal (Ra-T2). Time for removal procedures was also recorded. The third measurements were made after polishing (Ra-T3). Scanning Electronic Microscopy was performed in two samples of each group: after resin removal and after polishing. Results of roughness and time measurements were statically analysed by analysis of variance with post-hoc Bonferroni., Results: After polishing, tungsten carbide (P=0.1555) and fibreglass burs provided final surface roughness statistically similar to the baseline condition (P=1.0000). Yet, polymer burs, associated (P<0.0001) or not to alcohol (P<0.0001), provided surface roughness significantly higher when compared to baseline values. Polymer burs were more time-consuming on resinous remnant removal than tungsten carbide and fibreglass burs (P<0.05)., Conclusion: Polymer burs were less effective and more time-consuming to remove the remaining resin than tungsten carbide and fibreglass burs. The polishing step created smoother surfaces regardless of the burs used for resin removal., (Copyright © 2020 CEO. Published by Elsevier Masson SAS. All rights reserved.)

Low-power laser irradiation (LPLI) is clinically used to modulate inflammation, proliferation and apoptosis. However, its molecular mechanisms are still not fully understood. This study aimed to describe the effects of LPLI upon inflammatory, apoptotic and proliferation markers in submandibular salivary glands (SMGs) in an experimental model of chronic disorder, 24h after one time irradiation. Diabetes was induced in rats by the injection of streptozotocin. After 29 days, these animals were treated with LPLI in the SMG area, and euthanized 24h after this irradiation. Treatment with LPLI significantly decreased diabetes-induced high mobility group box 1 (HMGB1) and tumor necrosis factor alpha (TNF-α) expression, while enhancing the activation of the transcriptional factor cAMP response element binding (CREB) protein. LPLI also reduced the expression of bax, a mitochondrial apoptotic marker, favoring the cell survival. These findings suggest that LPLI can hamper the state of chronic inflammation and favor homeostasis in diabetic rats SMGs., Competing Interests: The authors have declared that no competing interests exist.

Basic dosimetric studies are necessary to support the use of photobiomodulation therapy (PBMT), since the great variety of laser parameters that are reported in the literature have created an obstacle to identifying reproducible results. Thus, the present study evaluates the process of tissue repair after the photobiomodulation therapy, taking into consideration the dose, frequency and the mode of energy delivery used. For this, 6 mm diameter wounds were created on dorsal skin of Wistar rats, and the animals were divided in control and irradiated groups, where L1 and L4 (irradiated with 1 point of 10 J/cm 2 ), L2 and L5 (5 points of 10 J/cm 2 ), L3 and L6 (1 point of 50 J/cm 2 ), respectively for one or multiple days of irradiations. A diode laser, λ 660 nm, 40 mW of power and 0.028 cm 2 of spot area was used. Our data showed that the group receiving multiple treatments over the first week post wounding, applied at 10 J/cm2 at each of 5 points on and around the wound (group L5) presented the best improvement of wound closure, higher cytokeratin 10, lower macrophage infiltration, and greater tissue resistance to rupture. We conclude that PBMT improves the skin wound healing process, and the outcomes were directly related to the chosen laser parameters and irradiation mode., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Salivary gland aquaporins (AQPs) are essential for the control of saliva production and maintenance of glandular structure. However, little is known of their role in salivary gland neoplasia. Salivary gland tumors comprise a heterogeneous group of lesions, featuring variable histological characteristics and diverse clinical behaviors. Mucoepidermoid carcinoma (MEC) is the most common salivary gland malignancy. The aim of this study was to evaluate the expression of AQP1, AQP3, and AQP5 in 24 MEC samples by immunohistochemistry. AQP1 expression was observed in vascular endothelium throughout the tumor stroma. AQP3 was expressed in epidermoid and mucosal cells and AQP5 was expressed in mucosal cells of MEC. These proteins were expressed in the human MEC cell line UH-HMC-3A. Cellular ultrastructural aspects were analyzed by electron microscopy to certificate the tumor cell phenotype. In summary, our results show that, despite the fact that these molecules are important for salivary gland physiology, they may not play a distinct role in tumorigenesis in MEC. Additionally, the in vitro model may offer new possibilities to further investigate mechanisms of these molecules in tumor biology and their real significance in prognosis and possible target therapies.

Objectives: To investigate the effects of irradiation lasers and light-emitting diode (LED) light on root resorption in rat molars during orthodontic tooth movement (OTM)., Materials and Methods: Twenty-one 12-week-old Wistar male rats were divided into three groups: OTM only (control [CG]), OTM and LED irradiation (DG), and OTM and low laser irradiation therapy (LG). The distance between the first and second molars was used to evaluate the amount of tooth movement. The mesial surfaces of the distopalatal roots were analyzed by scanning electron microscopy, and the area of the resorption was calculated., Results: Statistical analysis showed a decreased amount of tooth movement in the exposed DG or LG compared with CG, which was statistically significant ( P = .031 and P = .004, respectively). However, when the DG and LG groups were compared, no statistically significant differences were found ( P = .504). The root resorption areas were similar between CG and DG. However, statistically significant differences were found between LG and CG ( P = .014) and LG and DG ( P = .038)., Conclusions: Phototherapy did not enhance tooth movement, while infrared laser irradiation did increase root resorption.

This study describes the effect of the software binning and data averaging during micro CT volume acquisition, on the assessment of root resorption volumes. The mesial roots (n = 9), after orthodontic tooth movement during 14 days, were scanned, using a micro CT system (9 µm/pixel). All roots were reconstructed and the volumes of the resorption lacunae evaluated. The height and width of the pixels vary according to the parameters (A1, A2, A3, A4, A5, A6, A7, A8, A9) used during the scan. In the root #1 the mean volumes of resorption were similar in A4 and A7; in the root #2 there was no similarity in the mean volumes of resorption in any of the parameters; in root #3 only A4 presented mean volume different from zero (3.05 × 10°). In the root #5, the A1 and A7 presented similar mean volumes and in the A6 and A9 presented near mean volumes. In the root #9 the A1, A4, and A7 presented similar mean volumes and A6 and A9 also had similar mean volumes. Significant difference was detected in the volume of resorption among the roots #2, #5 and #9 (p = 0.04). When analyzing delicate structures such as the roots of rats' molars, the variation of such parameters will significantly influence the results.

This chapter describes laboratory protocols for TEM and SEM approaches allowing the examination of the dental hard tissues' constituents at the ultrastructural level. TEM has the highest resolution power to examine the cellular and extracellular matrix ultrastructure inside a given sample, detecting the presence, location, and quantification of organelles related to the metabolism of the cell type as well as membrane specializations. SEM allows the observation of the sample surface, for examining dimensional topography and distribution of exposed features.

Objective: Radiation-related caries are one the most undesired reactions manifested during or after head and neck radiotherapy. Fluoride application is an important strategy to reduce demineralization and enhance remineralizaton. To evaluate the effect of the topical application of fluoride during irradiation on dental enamel demineralization., Material and Methods: Thirty molars were randomly divided into three groups: Non-irradiated (NI), Irradiated (I), Irradiated with fluoride (IF). Each group was subdivided according to the presence or absence of pH-cycling (n=5). In the irradiated groups, the teeth received 70 Gy. The enamel's chemical composition was measured using Fourier Transform Infrared Spectrometry (organic matrix/mineral ratio - M/M and relative carbonate content - RCC). Vickers microhardness (VHN) and elastic modulus (E) were evaluated at three depths (surface, middle and deep enamel). Scanning electron microscopy (SEM) was used to assess the enamel's morphology., Results: The FTIR analysis (M/M and RCC) showed significant differences for irradiation, pH-cycling and the interaction between factors (p<0.001). Without pH-cycling, IF had the lowest organic matrix/mineral ratio and relative carbonate content. With pH-cycling, the organic matrix/mineral ratio increased and the relative carbonate content decreased, except for IF. VHN was influenced only by pH-cycling (p<0.001), which generated higher VHN values. ANOVA detected significant differences in E for irradiation (p<0.001), pH-cycling (p<0.001) and for the interaction between irradiation and pH-cycling (p<0.001). Increased E was found for group I without pH-cycling. With pH-cycling, groups I and IF were similar, and showed higher values than NI. The SEM images showed no morphological changes without pH-cycling. With pH-cycling, fluoride helped to maintain the outer enamel's morphology., Conclusions: Fluoride reduced mineral loss and maintained the outer morphology of irradiated and cycled enamel. However, it was not as effective in preserving the mechanical properties of enamel. Radiotherapy altered the enamel's elastic modulus and its chemical composition.

Objective:  The main purpose of this study was to evaluate the characteristics of a vegetal polymeric biomaterial intended for bone substitution in horses and to investigate the responses of the equine third metacarpal bone to biomaterial implantation., Materials and Methods:  Six horses were submitted to osteotomy on the dorsal aspect of the left and right third metacarpal bones; one bone defect was randomly selected for treatment with biopolymer, while the other was left untreated and served as a control. Bone density was monitored radiographically after surgery and bone biopsy fragments were collected at the end of the 120-day follow-up period. Biopsy fragments were analysed using light and scanning electron microscopy., Results:  Mean bone density values (mmAL) were greater in control defects (16.33 ± 1.6) than in polymer-treated defects (14.17 ± 1.7) at 120 days ( p  = 0.027). Light microscopy revealed greater percentages of new bone formation in control defects (50.15 ± 14.8) than in polymer treated defects (26.94 ± 12.1) at 120 days ( p  < 0.0001). Scanning electron microscopy analysis suggested a similar quality of pre-existing bone and new bone formed in the presence of biomaterial., Clinical Significance:  The absence of adverse reactions supports biomaterial biocompatibility and osteoconducting capacity and suggests the castor oil polymer is a suitable bone substitute for the treatment of bone defects in horses., Competing Interests: None., (Georg Thieme Verlag KG Stuttgart · New York.)

Objective: The purpose of this study was to histologically evaluate pulp and dentin under induced tooth movement (ITM) with different types of forces., Material and Methods: The maxillary right first molars of rats were submitted to movement with continuous (CF), continuous interrupted (CIF) and intermittent (IF) forces during 5, 7 and 9 days with nickel-titanium (NiTi) closed-coil springs exerting 50cN force magnitude. The groups were histologically evaluated as for cellularity pattern, presence of dystrophic, hemodynamic alterations in the pulp as well dentin alterations. The main observed alterations were related to hemodynamic pulp characteristics, such as presence of thrombosis, vascular congestion and hemorrhages. The hemodynamic alterations were statistically evaluated by Shapiro-Wilk normality test and analysis of variance by the Kruskall-Wallis test., Results: There was no significant differences observed between groups in the different types of applied forces and duration of ITM (vascular congestion, p=1.000; hemorrhage, p=0.305; thrombosis, p=1.000)., Conclusions: Pulp tissue alterations resulting from ITM were limited to hemodynamic events, without progressing to irreversible degeneration, regardless of the type of force applied.

Objective: Three-dimensional (3D) cell culture methods are of high importance to studies of biological processes. This is particularly the case with spheroid cultures, which create 3D cell aggregates without the use of exogenous materials. Compared to conventional monolayer cultures, cellular spheroid cultures have been demonstrated to improve multilineage potential and extracellular matrix production. To address this issue in depth, we present a more comprehensive analysis of 3D human dental pulp cell (hDPC) spheroids., Design: hDPC spheroids were fabricated by the pellet culture method and were cultured without adding any reagent to induce differentiation. The gene-expression profiles of the 3D and two-dimensional (2D) cultured hDPCs were compared by complementary DNA microarray analysis. Odontoblastic and osteoblastic differentiation marker gene expression was evaluated by quantitative real-time PCR (RT-qPCR). Hematoxylin-eosin (HE) staining and transmission electron microscopy (TEM) were applied to examine the morphology of hDPC spheroids and extracellular matrix components., Results: Compared with 2D monolayer culture, microarray analysis identified 405 genes and 279 genes with twofold or greater differential expression after 3 days and 28 days of 3D culture, respectively. In 3D hDPC spheroids, gene ontology analysis revealed upregulation of extracellular matrix-related genes and downregulation of cell growth-related genes. RT-qPCR analysis showed higher expression levels of osteocalcin, dentin sialophosphoprotein, and alkaline phosphatase. TEM revealed the morphological characteristics of the fibrillar collagen-rich matrix and cell-cell interactions., Conclusions: The present findings provide clues to understanding the mechanisms of pellet-cultured hDPCs and contribute to future research in the comparative studies of different 3D culture methods., (Copyright © 2018. Published by Elsevier Ltd.)

Objectives: This study describes the synthesis of brushite nanoparticles (CaHPO 4 ·2H 2 O) functionalized with triethylene glycol dimethacrylate (TEGDMA) and their application in dental restorative composites with remineralizing capabilities., Methods: Nanoparticles were synthesized, with TEGDMA being added to one of the precursor solutions at three different molar ratios (0:1, 0.5:1 and 1:1, in relation to the ammonium phosphate precursor). Then, they were added (10 vol%) to a photocurable dimethacrylate matrix containing 50 vol% of reinforcing glass particles. The resulting composites were tested for degree of conversion, biaxial flexural strength and elastic modulus (after 24h and 28days in water), and ion release (over a 28-day period). Commercial composites (one microhybrid and one microfilled) were tested as controls., Results: The final TEGDMA content in the functionalizing layer was modulated by the molar ratio added to the precursor solution. Functionalization reduced nanoparticle size, but did not reduce agglomeration. Improved mechanical properties were found for the composite containing nanoparticles with higher TEGDMA level in comparison to the composite containing non-functionalized nanoparticles or those with a low TEGDMA level. All brushite composites presented statistically significant reductions in strength after 28 days in water, but only the material with high-TEGDMA nanoparticles retained strength similar to the microhybrid commercial control. Overall, ion release was not affected by functionalization and presented steady levels for 28 days., Significance: Though agglomeration was not reduced by functionalization, the improvement in the matrix-nanoparticle interface allowed for a stronger material, without compromising its remineralizing potential., (Copyright © 2018 The Academy of Dental Materials. Published by Elsevier Inc. All rights reserved.)

Lipid droplet accumulation has been related to salivary gland hypofunction in diabetes. In this study, the effect of laser irradiation on the parotid glands (PGs) of diabetic rats was analyzed with regard to its effect on lipid droplet accumulation, intracellular calcium concentration and calmodulin expression. The animals were distributed into 6 groups: D0, D5, D20 and C0, C5, C20, for diabetic (D) and control animals (C), respectively. Twenty-nine days following diabetes induction, PGs of groups D5 and C5; D20 and C20 were irradiated with 5 and 20 J/cm 2 of a red diode laser at 100 mW, respectively. After 24 hours, PGs were removed for histological, biochemical, and western blotting analysis. The diabetic animals showed lipid droplet accumulation, which was decreased after irradiation. Ultrastructurally, the droplets were nonmembrane bound and appeared irregularly located in the cytoplasm. Moreover, diabetic animals showed an increased intracellular calcium concentration. In contrast, after laser irradiation a progressive decrease in the concentration of this ion was observed, which would be in agreement with the results found in the increased expression of calmodulin in D20. These data are promising for using laser to decrease lipid droplet accumulation in PGs, however, more studies are necessary to better understand its mechanisms. Micrographs showing decreased lipid accumulation after laser irradiation in light micrographs (LM), and morphology of lipid droplet in transmission electron microscopic (TEM). LM: (A) PGs from nondiabetic rats that did not receive Laser irradiation (LI), (B) PGs from nondiabetic rats that received a dose of 20 J/cm 2 , (C) lipid accumulation (arrows) in the secretory cells from diabetic rats that did not receive irradiation, (D) reduction of lipid accumulation in the secretory cells from diabetic rats that received a dose of 20 J/cm 2 and TEM: (E) scale bar = 5 μm, (F) scale bar = 1 μm, and (G) scale bar = 0.5 μm., (© 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Objectives: The objective of this study was to evaluate the biomechanical properties of dentin and the microtensile bond strength (μTBS) performed before or after radiotherapy (RT)., Material and Methods: Dentin chemical composition (infrared spectroscopy-FTIR), SEM images, and mechanical properties (Vickers microhardness-VHN and elastic modulus-E) were evaluated comparing no irradiated and irradiate dentin (n = 5). RT was defined by application of 72 Gy (1.8 Gy daily, 5 days per week, during 8 weeks) with sample immersed in distilled water. μTBS evaluated three groups (n = 10): NI-no irradiated; IB-irradiation before restoration; and IA-irradiation after restoration. Resin-dentin sticks (1.0 mm 2 ) were obtained and submitted to μTBS. Analysis of the bonding interface was made by confocal microscopy., Results: After RT, percentage ratio of FTIR analysis showed increased absorption for all bands. SEM image showed a disorganized dentin structure. Two-way ANOVA showed increased VHN (p = 0.005) and decreased E (p < 0.001). For μTBS, one-way ANOVA and Duncan test showed significant differences among groups (p = 0.018). IB group presented the lowest bond strength values., Conclusions: RT alters the absorption bands and SEM images showed a disorganization of the dentin structure. Mechanical properties were changed with increased VHN and decreased E. μTBS was affected by the radiotherapy and restoration period (before or after)., Clinical Relevance: RT causes changes that contribute to increased risk of tooth decay. Restorative treatments can be performed using adhesive procedures, but it is preferable to be performed before of the irradiation protocol, to guarantee better adhesive properties to restoration.

Tissue bioengineering has been applied to Endodontics to seek a more biological treatment. The presence of blood vessels is crucial for cell nutrition during tissue formation. Objective This study analysed the application of vascular endothelial growth factor (VEGF) in the angiogenesis of mature root canals. Material and methods Upper first molars of twelve 13-week old Wistar male rats were used. The root pulp of the mesiobuccal canal was removed and the root canal instrumented with K-files up to size #25. Periapical bleeding was induced into the root canal by introducing a #15 K-file beyond the apex. The teeth on the right side of the arch were filled up with blood clot (G1), whereas those on the left side were filled up with blood clot plus 50 ng/ml of VEGF (G2). Teeth were sealed with light-curing glass-ionomer cement and the animals were sacrificed after 60 days. The maxilla was dissected and fixed before obtaining serial sections for histological processing with haematoxylin-eosin (HE) and immunohistochemical factor-VIII. Immunohistochemical labelling was evaluated using scores for statistical analysis. Results Immunohistological analysis demonstrated the presence of angiogenesis in both groups, but with higher angiogenic maturation in G2 during the experimental period (p<0.05). HE staining showed connective tissue with absence of odontoblasts in all specimens. Conclusions It can be concluded that it is possible to obtain angiogenesis in mature root canals with or without the use of VEGF, although the latter tends to accelerate blood vessel formation.

OBJECTIVE To evaluate the efficacy of castor oil polyurethane polymer with calcium carbonate for use in a unicortical ostectomy on the dorsal surface of the third metacarpal bone of horses. ANIMALS 6 adult horses. PROCEDURES A unicortical ostectomy was created on the dorsal surface of both third metacarpal bones of each horse. Castor bean (Ricinus communis) oil polyurethane polymer with calcium carbonate was implanted into the ostectomy on 1 limb, and the ostectomy of the contralateral limb was left unfilled and served as a control sample. Ostectomy sites were evaluated histologically 120 days later. Biopsy specimens were obtained from the interface of bone and polymer or the interface of bone and newly formed tissue; specimens were processed for histomorphometric evaluation by use of light microscopy, immunohistochemical analysis, histochemical analysis, and transmission electron microscopy. RESULTS Osteoconductive activity of the biomaterial was confirmed by the presence of osteoblasts in the biopsy specimens. Absence of a chronic inflammatory response or foreign body reaction indicated biocompatibility. Expression of osteoblast markers was detected in the newly formed tissue. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that castor oil polyurethane polymer with calcium carbonate could be an acceptable compound for use as a bone substitute in horses with fractures in which bone filling is necessary.

Silver phosphate is a semi-conductor sensitive to UV-Vis radiation (<530nm). Exposure to radiation removes electrons from the oxygen valence shell, which are scavenged by silver cations (Ag + ), forming metallic silver (Ag 0 ) nanoparticles. The possibility of silver nanoparticle formation in situ by a photoreduction process was the basis for the application of mixed calcium phosphate/silver phosphate particles as remineralizing and antibacterial fillers in resin-based dental materials. Mixed phosphate particles were synthesized, characterized and added to a dimethacrylate resin in 20% or 30% mass fractions to investigate their efficacy as ion-releasing fillers for dental remineralization and antibacterial activity. The formation of metallic silver nanoparticles after exposure to visible radiation from a dental curing unit (peak emission: 470nm) was demonstrated by particle X-ray diffraction and scanning electron microscopy analysis of the composite fractured surface. Calcium and phosphate release from materials containing the mixed particles were similar to those containing pure CaP particles, whereas Streptococcus mutans colonies were reduced by three orders of magnitude in relation to the control, which can be attributed to silver release. As expected, the optical properties of the materials containing mixed phosphate particles were compromised by the presence of silver. Nevertheless, materials containing mixed phosphate particles presented higher fracture strength and elastic modulus than those with pure CaP particles., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Osteoclasts and chondroclasts are necessary, during endochondral ossification, for the resorption of primary bone and calcified cartilage septa, respectively. The bisphosphonates inhibit mineralized tissue resorption by various mechanisms according to the different types of this drug, which can affect bone remodeling during skeletal growth. The objective of the present study is to analyze the way that alendronate (ALN) and etidronate (ETN) can affect osteoclastogenesis and bone formation during endochondral ossification of the long bones of growing rats. Newborn Wistar rats were treated daily with ETN, ALN, or sterile saline solution (control) for 21 days. Their femur and tibiae epiphyses were radiographed and analyzed by light, scanning and transmission electron microscopy. The expression of genes related to osteogenesis and to osteoclast differentiation and activity were analyzed by real-time quantitative polymerase chain reaction. The ETN group presented reduced body weight, disorganized growth plate and an extended area of cartilage in the ossification zone with little bone matrix; in the ALN group, this area was not altered. The ALN presented latent TRAP-positive cells, whereas in the ETN group, they were activated. The expression of NFκB1 and 2, OPG, Spp1 and Runx2 in the ossification zone was reduced by both bisphosphonates. RANKL expression was reduced by ETN, whereas ALN decreased the expression of RANK. The results also indicated that, in addition to the anti-resorptive effect of the drugs, disturbances in bone deposition occurred concomitantly with the reduced expression of osteogenesis-related genes.

Diabetes can lead to dysfunction of the secretory capacity in salivary glands. Activation of the receptor for advanced glycation end products (RAGE) and its ligands has been suggested to participate in chronic disorders such as diabetes and its complications. In this study, the expression of RAGE, high mobility group box 1 (HMGB1) and advanced glycation end products (AGE), as well as the effects of low-power laser irradiation (LPLI) in salivary glands of diabetic rats were evaluated, and the mechanisms involved were characterized. The expression of RAGE and HMGB1 at the protein and mRNA levels was observed in submandibular glands (SMGs) of streptozotocin-induced diabetic rats. A diode laser was applied at 660 nm, 70 mW, 20 J/cm2, 0.56 J/point, with a spot area of 0.028 cm2 and its in vivo effects and the pathways involved were evaluated. Immunohistochemistry and western blotting analysis were performed for inflammatory and apoptosis markers. Diabetes up-regulates HMGB1/AGE/RAGE axis gene expression in SMGs that is associated with activation of the nuclear factor kappa B (NF-κB) pathway. Interestingly, LPLI suppresses NF-κB activation induced by inflammation. LPLI also reduces diabetes-induced apoptosis. That effect was accompanied by decreased levels of Bax, and cleaved caspase 3, which were up-regulated in diabetes. Taken together, our data suggest that LPLI reduces diabetes-induced inflammation by reducing the induction of HMGB1, ultimately leading to inhibition of apoptosis in submandibular glands of diabetic rats., Competing Interests: The authors have declared that no competing interests exist.

Dicalcium phosphate dihydrate particles functionalized with triethyleneglycol dimethacrylate were synthesized and added to a photocurable mixture of bisphenol-A glycidyl dimethacrylate and triethyleneglycol dimethacrylate with the purpose of developing a resin composite capable of releasing calcium and phosphate ions to foster dental remineralization. Particle functionalization would minimize the deleterious effect of adding low cohesive strength nano-structured particles with no chemical interaction with the organic matrix on the material's mechanical properties. The results showed that calcium release over 28 days was not impaired by particle functionalization. A statistically significant 32% increase in strength was recorded with the use of functionalized dicalcium phosphate dihydrate in comparison to the material containing non-functionalized particles. However, the strength of the unfilled resin was not matched by the composite with functionalized particles. Elastic modulus increased with particle incorporation, regardless of functionalization. Degree of conversion and optical properties (total transmittance and color change/ΔE) of the resin-based materials were not affected by the addition of dicalcium phosphate dihydrate particles (functionalized or not).

Oral mucositis (OM) is a debilitating consequence of cancer treatment that could be treated with photobiomodulation therapy (PBMT); however, there is no consensus about its dosimetric parameters for OM healing. The aim of this study was to compare different PBMT protocols on OM treatment, through clinical and histological analysis. Thirty hamsters were used, in an induced model of OM by 5-fluorouracil (5-FU) and superficial scratching, in seven days of follow-up. The animals were divided into five groups: control (C), which received only anesthesia and chemotherapeutic vehicle; chemotherapy (Ch), which received anesthesia, 5-FU, and scratches; laser 1 (L1), the same as Ch group, PBMT 6 ?? J / c m 2 and 0.24 J (one point); laser 2 (L2), the same as Ch group, PBMT 25 ?? J / c m 2 and 1 J (one point); and laser 3 (L3), the same as Ch group, PBMT 4 points of 0.24 J and 6 ?? J / c m 2 each. The laser used has ? = 660 ?? nm , 0.04 ?? c m 2 of spot area, and 40 mW. The best PBMT protocol to maintain lowest OM levels compared to Ch group was L1, followed by L2 and L3. Our results suggest that the application mode of PBMT and the energy delivered per area could interfere with the OM healing.

Objective: To evaluate the strength and ion release of experimental composites containing TEGDMA-functionalized calcium phosphate particles., Methods: Seven composites containing equal parts (in mols) of BisGMA and TEGDMA and 60vol% of fillers were manipulated. Filler phase was constituted by silanized barium glass and 0% (control), 10% or 20% (volume) of dicalcium phosphate dihydrate (DPCD) particles, either non-functionalized or functionalized with two different TEDGMA contents. DCPD particles were synthesized and characterized by X-ray diffraction (XRD), elemental analysis, surface area and dynamic light scattering. Composites were tested for degree of conversion (DC) by near-FTIR. Biaxial flexural strength (BFS) was determined after 24h and 28days in water. Calcium and phosphate release after 7days was assessed using inductively coupled plasma optical emission spectrometry (ICP-OES). Data were analyzed by ANOVA/Tukey test (alpha:5%)., Results: XRD confirmed the crystalline structure corresponding to DCPD. Elemental analysis revealed particles with zero, 14% or 22% TEGDMA, with similar D 50 (around 19μm) and surface areas from 3.5 to 11.4m 2 /g. The presence of DCPD did not reduce DC. After 24h, functionalization (both 14% and 22% TEGDMA) improved composite strength in comparison to non-functionalized DCPD, both at 10% and 20% levels. After 28days, BFS of materials containing 10% functionalized DCPD were statistically similar to the control containing only barium glass. Among composites containing 10% DCPD, particle functionalization with 14% TEGDMA did not jeopardize ion release., Significance: At 10vol%, the use of TEGDMA-functionalized CaP particles improved composite strength in relation to non-functionalized particles, while maintaining similar ion release levels., (Copyright © 2016 The Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.)

In this study, clinical, biochemical and histological analysis were used to compare different phototherapies, including LED, low and high-power laser (HPL) for the treatment of chemotherapy (CT)-induced oral mucositis (OM). One-hundred-fifty hamsters were divided into five groups: C: control; CH: CT/OM induction; L: CT/OM induction and treatment with LED (635 nm, 1.2 J), HL: CT/OM induction and treatment with HPL (808 nm, 10 J), LL: CT/OM induction and treatment with low-level laser therapy (LLLT) (660 nm, 1.2 J). OM was induced by scratches performed on check pouch mucosa after two injections of 5-Fluorouracil. The experiment lasted 10 days and OM was analyzed by specific clinical scales on days 5, 7 and 10. The animals were euthanized and the cheek pouch mucosa removed for biochemical (TNF-α concentration) and histological (light microscopy) analysis. After statistical analysis, the authors' results showed LED and LLLT therapies were efficient treatments for OM, decreasing TNF-α concentration on day 7 (p < 0.05) and completely healing the mucosa on day 10. HPL showed no interference in final healing of OM. According to the methodology used and the results obtained in the present study, LLLT and LED therapies were the best choices to decrease the severity of OM, accelerating tissue repair and decreasing the inflammatory process. Clinical evaluation of OM in Groups CH, LL, L and HL and their respective arrangement of phototherapy treatments at different time intervals (5, 7 and 10 days)., (© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

In this study, we have analyzed the viability and cell growth, as well as, the mineralization of extracellular matrix (ECM) by alizarin red and von Kossa staining of calvaria-derived osteogenic cultures, treated with TGF-β1 alone or associated with Dex comparing with acid ascorbic (AA) + β-glicerophosphate (βGP) (positive mineralization control). The expression of the noncollagenous proteins bone sialoprotein (BSP), osteopontin (OPN) and fibronectin (FN) were evaluated by indirect immunofluorescence. In addition, the main ultrastructural morphological findings were assessed by transmission electron microscopy. Osteogenic cells were isolated of calvaria bone from newborn (2-day-old) Wistar rats were treated with TGF-β1 alone or with dexamethasone for 7, 10, and 14 days. As positive mineralization control, the cells were supplemented only with AA+ βGP. As negative control, the cells were cultured with basal medium (α-MEM + 10%FBS + 1%gentamicin). The treatment with TGF-β1, even when combined with Dex, decreased the viability and cell growth when compared with the positive control. Osteoblastic cell cultures were positive to alizarin red and von Kossa stainings after AA + βGP and Dex alone treatments. Positive immunoreaction was found for BSP, OPN and FN in all studied treatments. Otherwise, when the cell cultures were supplemented with TGF-β1 and TGF-β1 + Dex, no mineralization was observed in any of the studied periods. These present findings suggest that TGF-β1, in the studied in vitro doses, inhibits the proliferation and differentiation of osteoblastic cells by impairment of nodule formation., (© 2016 Wiley Periodicals, Inc.)

Introduction: Studies defining the characteristics of light curing units and photoactivation methods are necessary to allow the correct choices to be made in daily practice. This study aimed to determine whether different photoactivation protocols for composite resins [periodic level shifting (PLS) - 5 second and soft-start] are able to maintain or enhance the mechanical properties and marginal adaptation of restorations., Methods: Restorations were placed in bovine teeth using the following photoactivation methods: continuous light for 20 seconds (control group); PLS technology (PLS - 5 second group); and continuous light and a light guide tip distance of 6 mm after which the tip was placed at the surface of the restoration (soft-start group). The teeth were transversely sectioned in the incisal-cervical direction. Thirty halves were randomly selected for Knoop microhardness testing (n = 10). The other 30 halves were subjected to scanning electron microscopy analysis. The images obtained were measured to identify the highest marginal gap, and statistical tests for variance analysis were conducted., Results: Microhardness tests showed no statistically significant difference between the photoactivation methods analysed (P ≥ 0.01). The tests showed a difference among depths (P < 0.01), with the deeper layers being the hardest. In analysing marginal adaptation, no significant difference was identified between the higher marginal gap values in the continuous (mean = 10.36) and PLS - 5 second (mean = 10.62) groups, and the soft-start group (mean = 5.83) presented the lowest values (P < 0.01)., Conclusions: The PLS - 5 second and soft-start protocols did not alter the hardness of the restorations. Moreover, the PLS - 5 second protocol did not alter the marginal adaptation, whereas the soft-start protocol improved marginal adaptation., (© 2016 FDI World Dental Federation.)

Whole genome sequencing and analyses of Ureaplasma diversum ATCC 49782 was undertaken as a step towards understanding U. diversum biology and pathogenicity. The complete genome showed 973,501 bp in a single circular chromosome, with 28.2% of G+C content. A total of 782 coding DNA sequences (CDSs), and 6 rRNA and 32 tRNA genes were predicted and annotated. The metabolic pathways are identical to other human ureaplasmas, including the production of ATP via hydrolysis of the urea. Genes related to pathogenicity, such as urease, phospholipase, hemolysin, and a Mycoplasma Ig binding protein (MIB)-Mycoplasma Ig protease (MIP) system were identified. More interestingly, a large number of genes (n = 40) encoding surface molecules were annotated in the genome (lipoproteins, multiple-banded antigen like protein, membrane nuclease lipoprotein and variable surface antigens lipoprotein). In addition, a gene encoding glycosyltransferase was also found. This enzyme has been associated with the production of capsule in mycoplasmas and ureaplasma. We then sought to detect the presence of a capsule in this organism. A polysaccharide capsule from 11 to 17 nm of U. diversum was observed trough electron microscopy and using specific dyes. This structure contained arabinose, xylose, mannose, galactose and glucose. In order to understand the inflammatory response against these surface molecules, we evaluated the response of murine macrophages J774 against viable and non-viable U. diversum. As with viable bacteria, non-viable bacteria were capable of promoting a significant inflammatory response by activation of Toll like receptor 2 (TLR2), indicating that surface molecules are important for the activation of inflammatory response. Furthermore, a cascade of genes related to the inflammasome pathway of macrophages was also up-regulated during infection with viable organisms when compared to non-infected cells. In conclusion, U. diversum has a typical ureaplasma genome and metabolism, and its surface molecules, including the identified capsular material, represent major components of the organism immunopathogenesis., Competing Interests: The authors have declared that no competing interests exist.

This study was conducted to evaluate the effects of treatment with strontium ranelate (SR) on the repair of bone defects and molecular components of bones in femurs. Adult female rats (n=27) were subjected to ovariectomy (OVX) or Sham surgery. Thirty days after surgery, a defect was made in the femur and the animals were then divided into three groups: OVX, SHAM and OVX+SR. Euthanasia was performed four weeks after the bone defect surgery. Repair in bone defect was assessed by computed microtomography (μCT) and chemical composition of cortical bone was analyzed by Fourier transform infrared (FTIR) spectroscopy and energy dispersive X-ray spectroscopy (EDS). The trabecular thickness (Tb.Th) of the newly formed bone in the OVX+SR group was significantly higher than that for the OVX group. The collagen maturity in the OVX+SR group was smaller than in the other two groups. In this group, a significant increase in the amount of strontium (Sr) and a decrease in the amount of calcium (Ca) embedded to bone tissue were also observed. Systemic treatment with SR improved microarchitecture of the newly formed bone inside the defect, but decreased cross-linking of mature collagen in cortical bone.